Golpayegani Abdolali, Douraghi Masoumeh, Rezaei Farhad, Alimohammadi Mahmood, Nodehi Ramin Nabizadeh
1Department of Environmental Health Engineering, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Vice-Chancellor for Health, Bam University of Medical Sciences, Bam, Iran.
J Environ Health Sci Eng. 2019 Mar 7;17(1):407-416. doi: 10.1007/s40201-019-00359-w. eCollection 2019 Jun.
Lack of culturability in the viable but non-culturable (VBNC) bacteria and the ability to regain infectivity in favourable conditions is one of the new challenges of public health providers for monitoring in environmental samples. Propidium monoazide quantitative polymerase chain reaction (PMA-qPCR) is one of the promising methods for timely detection of VBNC pathogens in environmental samples. We developed and used a method for the first time to detection of VBNC in swimming pool water samples using a membrane filter (MF). Moreover, the dominant model of the distribution of colonies on the MF and the effect of the culture medium and MF type on colony recovery by MF were evaluated. Swimming pool samples were subjected to conventional culture-based, qPCR and PMA-qPCR methods and the results were compared for the presence of VBNC in the samples. The positivity rate was 21% and 75% for in water samples as confirmed by standard culture-based and qPCR methods, respectively. Furthermore, of 24 samples, 9 (37.5%) were positive for VBNC . The developed qPCR/PMA-qPCR assay can detect the VBNC bacteria directly from aquatic samples and may result in better monitoring of recreational waters.
活的非可培养（VBNC）细菌缺乏可培养性以及在有利条件下恢复感染性的能力是公共卫生机构在环境样本监测方面面临的新挑战之一。单叠氮碘化丙啶定量聚合酶链反应（PMA-qPCR）是及时检测环境样本中VBNC病原体的一种有前景的方法。我们首次开发并使用了一种利用膜过滤器（MF）检测游泳池水样中VBNC的方法。此外，还评估了MF上菌落分布的主导模型以及培养基和MF类型对MF菌落回收率的影响。对游泳池样本采用基于传统培养的方法、qPCR和PMA-qPCR方法，并比较结果以检测样本中VBNC的存在情况。基于标准培养的方法和qPCR方法分别确认水样中的阳性率为21%和75%。此外，在24个样本中，9个（37.5%）VBNC呈阳性。所开发的qPCR/PMA-qPCR检测方法可直接从水样中检测VBNC细菌，可能会更好地监测娱乐用水。
